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pgd1, an Arabidopsis thaliana deletion mutant, is defective in pollen germination

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Abstract

An Arabidopsis deletion mutant was fortuitously identified from the alpha population of T-DNA insertional mutants generated at the University of Wisconsin Arabidopsis Knockout Facility. Segregation and reciprocal crosses indicated that the mutant was a gametophytic pollen sterile mutant. Pollen carrying the mutation has the unusual phenotype that it is viable, but cannot germinate. Thus, the mutant was named pollen germination defective mutant 1 (pgd1), based on the pollen phenotype. Flanking sequences of the T-DNA insertion in the pgd1 mutant were identified by thermal asymmetric interlaced (TAIL) PCR. Sequencing of bands from TAIL PCR revealed that the T-DNA was linked to the gene XLG1, At2g23460, at its downstream end, while directly upstream of the T-DNA was a region between At2g22830 and At2g22840, which was 65 genes upstream of XLG1. Southern blotting and genomic PCR confirmed that the 65 genes plus part of XLG1 were deleted in the pgd1 mutant. A 9,177 bp genomic sequence containing the XLG1 gene and upstream and downstream intergenic regions could not rescue the pgd1 pollen phenotype. One or more genes from the deleted region were presumably responsible for the pollen germination defect observed in the pgd1 mutant. Because relatively few mutations have been identified that affect pollen germination independent of any effect on pollen viability, this mutant line provides a new tool for identification of genes specifically involved in this phase of the reproductive cycle.

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Acknowledgments

We thank Ms. Anne Gibson for technical assistance, Drs. Dazhong Zhao for help with pollen staining, Annick Stintzi and John Browse for suggestions on in vitro pollen germination experiment, Rosemary Walsh for help with SEM, Tom Jack for providing vector pD991, Mark Johnson for discussion on mutant pollen characterization, and Hong Ma, Sheila McCormick, Zhenbiao Yang, and Andy McCubbin for discussions on the project. This research was supported by United States Department of Agriculture grants 2001-35304-09916 and 2003-35304-13924 to S. M. A. S.M.A. also gratefully acknowledges support from a National Science Foundation POWRE grant (MCB-9973546), which enabled the initiation of this research during a sabbatical in the laboratory of Dr. Michael R. Sussman.

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  1. Liu-Min Fan

    Present address: Peking-Yale Joint Center for Plant Molecular Genetics and Agro-Biotechnology, National Laboratory of Protein Engineering and Plant Genetic Engineering, College of Life Sciences, Peking University, Beijing, 100871, China

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  1. Department of Biology, Penn State University, 208 Mueller Laboratory, University Park, PA, 16802, USA

    Lei Ding, Liu-Min Fan & Sarah M. Assmann

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  1. Lei Ding
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Correspondence to Sarah M. Assmann.

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Communicated by José Feijó.

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Ding, L., Fan, LM. & Assmann, S.M. pgd1, an Arabidopsis thaliana deletion mutant, is defective in pollen germination. Sex Plant Reprod 20, 137–149 (2007). https://doi.org/10.1007/s00497-007-0050-z

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